Method of lowering blood cholesterol level

ABSTRACT

The level of blood cholesterol in a living animal body in which a state of hypercholesterolemia exists is lowered by orally administering to said animal body a pharmaceutically effective amount of a polymer, which is (1) polymerized unsaturated carboxylic acid or anhydride, or (2) a copolymer of (a) an unsaturated monomer having, for Example, 2 to 30 carbon atoms and, (b) an unsaturated carboxylic acid, anhydride or derivative thereof as exemplified by octadecene-1/maleic anhydride copolymer. Polymers having lipophilic properties are a preferred class. A typical dosage is an amount which represents in the range of from about 0.01 to about 5.0% of the diet.

REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of copending application Ser.No. 188,577, filed Oct. 12, 1971 and now abandoned, which application isin turn a continuation-in-part of copending application Ser. No.789,081, filed Jan. 2, 1969, now abandoned.

BACKGROUND OF THE INVENTION

This invention relates to a method for lowering the blood cholesterollevel in a living animal body, in which a state of hypercholesterolemiaexists. In a particular aspect this invention relates to a method forlowering the blood cholesterol level in a living animal body in which astate of hypercholesterolemia exists by orally administering to theliving animal body a pharmaceutically effective amount of a polymerselected from the group consisting of (1) polymerized unsaturatedcarboxylic acid or anhydride and (2) a copolymer of (a) an unsaturatedmonomer having, for example, 2 to 30 carbon atoms, such as, alkene,alkylcarboxyalkene, phenylalkene and alkoxyalkene and, (b) anunsaturated carboxylic acid, anhydride or derivative thereof.

The link between high levels of blood cholesterol (hypercholesterolemia)and cardio-vascular disease in warmblooded vertebrates is wellestablished. The malady is known not only to contribute to chronicconditions of high blood pressure, but to increase the risk ofincapacitating and often fatal coronary attacks.

SUMMARY OF THE INVENTION

It is an object of the present invention to provide a method forlowering the blood cholesterol in a living animal body in which a stateof hypercholesterolemia exists.

It is a further object of the present invention to provide compositionswhich are effective in the aforesaid method in lowering bloodcholesterol levels in a living animal body in which a state ofhypercholesterolemia exists.

Other objects and advantages of the present invention will be apparentfrom the specification and appended claims.

The present invention resides in the discovery that the level of bloodcholesterol in a living animal body in which a state ofhypercholesterolemia exists is lowered by orally administering to saidanimal body a pharmaceutically effective amount of a polymer selectedfrom the group consisting of (1) a polymerized unsaturated carboxylicacid, or anhydride (2) a copolymer of (a) an unsaturated monomer having,for example, 2 to 30 carbon atoms, and (b) an unsaturated carboxylicacid, anhydride or derivative thereof. By this method the bloodcholesterol level in living animal bodies, including warm-bloodedvertebrate animals, such as chickens, dogs, cats, cattle, swine andprimates, for example monkeys, is effectively lowered.

DETAILED DESCRIPTION

The polymer can be orally administered to the living animal body by anysuitable means, and in any suitable form. For example, the polymer canbe incorporated into ordinary foodstuffs and beverages containingnutritional values in an amount sufficient to produce the desiredreduction of blood cholesterol. Also, the polymer can be incorporatedinto a pharmaceutical composition of the form customarily employed fororal administration. Pharmaceutical compositions containing the polymermay be in liquid form, for example, a solution or suspensionspecifically adapted for oral administration or in solid form, forexample, a tablet, capsule, pill or packaged powder. Advantageously, thepharmaceutical composition containing the polymer can be prepared inunit dosage form using pharmaceutically acceptable carriers, such as,for example, starch, glucose, lactose, gelatin, sucrose, etc. and thelike. If desired, the dosage unit can be made up in a sustained releaseform to give a controlled dosage over an extended period of time.

The amount or dosage of polymer administered to the living animal bodywill, of course, vary depending among other things, on the size of theliving animal body, the particular living animal body to be treated, thelevel of blood cholesterol, and the general health of the living animalbody, and any pharmaceutically effective amount may be employed. Thedosage can be determined with regard to established medical practice.Generally, the amount of polymer administered on a daily basis is in therange of from about 0.01 to about 5.0% of the total diet, and typicallyin the range of from about 0.05 to about 3.0%.

The polymer of use in the present invention may be water soluble orwater-insoluble. Many of the normally water-soluble polymers areconverted to the water-insoluble form by introduction of sufficientcrosslinks in the known manner. Crosslinking may be accomplished eitherduring the preparation of the polymer or by subsequent treatment of thepolymer to make the polymer insoluble in water. The water insolubilityof the polymer can be varied by regulation of the degree of crosslinkingof the polymer. The term "water-insoluble" as used herein is taken tomean that the polymer concerned does not dissolve in water or aqueoussolutions, even though it may have such characteristics as a high degreeof swelling, due to solvation by water even to the extent of existencein a gel form. Such characteristics are typically imparted bycrosslinking.

As previously indicated, the polymers employed in the method of thepresent invention, are (1) polymerized unsaturated carboxylic acids oranhydrides, (2) copolymers of (a) an unsaturated monomer having, forexample, 2 to 30 carbon atoms, and (b) an unsaturated carboxylic acidanhydride or unsaturated carboxylic acid derivative, preferably having aweight average molecular weight of at least 1,000 and a degree ofpolymerization of at least 8.

The polymer may advantageously be an EMA-type polymer.

Among the EMA-type polymers suitable for the practice of the instantinvention are polymers and pharmaceutically acceptable salts of polymershaving units of the formula ##STR1## wherein: R_(A) and R_(B) areselected from the group consisting of hydrogen, halogen (preferablychlorine), alkyl of 1 to 4 carbon atoms (preferably methyl), cyano,phenyl, or mixtures thereof; provided that not more than one of R_(A)and R_(B) is phenyl; Z is a bivalent radical (preferably alkylene,phenylalkylene, alkoxyalkylene, alkylcarboxyalkylene and aliphaticacyloxyalkylene) of 1 to 30 carbon atoms, q is zero or one, X and Y areselected from hydroxy, --O alkali metal, OR, --OH--NH₃, --OH--R₃ N,--OH--R₂ NH, --OH--RNH₂, --NRR', --(Q)_(p) --W--(NR'R')_(x) and--(Q)_(p) --W--(-OH)_(x), wherein x is 1 to 4 and p is zero or one,wherein R is selected from the group consisting of hydrogen, alkyl,phenylalkyl, or phenyl, in each case of 1 to 18 carbon atoms, wherein R'is H or R, wherein Q is oxygen or --NR'--, and wherein W is a bivalentradical preferably selected from alkylene, phenylene, alkylene amine andphenylalkylene having up to 20 carbon atoms, X and Y taken together canbe oxygen or --NR--, --N--W-- (NR'R')_(x) or --N--W--(NR'R'R")_(x)+wherein R, W, R' have the meanings previously assigned and R" is alkylof 1 to 18 carbon atoms, benzyl or aromatic-substituted benzyl. Theunits of the formula given above are recurring n being at least 8 andcan be as much as 100,000 degrees of polymerization. When the units arerecurring the symbols in the various recurring units do not necessarilystand for the same thing in all of the recurring units.

Many of these polymers suitable for the practice of the presentinvention or suitable after conversion to derivatives are commerciallyavailable.

The polycarboxylic acid polymers can be of the nonvicinal-type includingthose containing monomer units, such as acrylic acid, acrylic anhydride,methacrylic acid, crotonic acid or their respective derivatives,including partial salts, amides and esters or of the vicinal type,including maleic, itaconic, citraconic, a-dimethyl maleic, a-butylmaleic, a-phenyl maleic, fumaric, aconitic, a-chloromaleic,a-bromomaleic, a-cyanomaleic acids including their salts, amides andesters. Anhydrides of the foregoing acids are also advantageouslyemployed.

Co-monomers suitable for use with the above polycarboxylic acid monomersinclude a-olefins, such as ethylene, 2-methyl-pentene-1, propylene,butylene, 1- or 2-butene, 1-hexene, 1-octene, 1-decene, 1-dodecene,1-octadecene, and other vinyl monomers, such as styrene, a-methylstyrene, vinyltoluene, vinyl acetate, vinyl chloride, vinyl formate,vinyl alkyl ethers, e.g. methylvinylether, alkyl acrylates, alkylmethacrylates, acrylamides, methacrylamides, alkylmethacrylamides andalkylacrylamides, or mixtures of these monomers. Reactivity of somefunctional groups in the copolymers resulting from some of thesemonomers permits formation of other useful functional groups in theformed copolymer, including hydroxy, lactone, amine and lactam groups.

Any of the said carboxylic acids or derivatives, may be copolymerizedwith any of the other monomers described above, and any other monomerwhich forms a copolymer with carboxylic acids or derivatives. Althoughthese copolymers can be prepared by direct polymerization of the variousmonomers, frequently they are more easily prepared by an after-reactionmodification of an existing copolymer. Copolymers are convenientlyidentified in terms of their monomeric constituents. The names soapplied refer to the molecular structure and are not limited to thepolymers prepared by the copolymerization of the specified monomers.

The initial copolymers of anhydrides and another monomer can beconverted to carboxyl-containing copolymers by reaction with water, andto ammonium, alkali and alkaline earth metal and alkylamine saltsthereof by reaction with alkali metal compounds, alkaline earth metalcompounds, amines or ammonia. Other suitable derivatives of the abovepolymers include the alkyl or other esters and amides, alkyl amides,dialkyl amides, phenylalkyl amides or phenyl amides prepared by reactingcarboxyl groups on the polymer chain with the selected amines or alkylor phenylalkyl alcohol, as well as amino esters, amino amides, hydroxyamides and hydroxy esters, wherein the functional groups are separatedby alkylene, phenyl, alkylene amine, alkylene oxide, phenylalkyl,phenylalkylphenyl, or alkylphenylalkyl or other aryl groups. Moietiesbearing amine or amine salts including quaternary salt groups areconveniently formed by reaction of the carboxyls of their anhydrideprecursors, where applicable with polyfunctional amines such asdimethylaminopropylamine or dialkylaminoalcohols such asdimethylaminoethanol, the former forming an amide linkage with thepolymer, or in certain cases at higher temperatures forming an imidelinkage with vicinal carboxyls, and the latter forming an ester linkage.Such pendant free amine groups can then be converted, if desired, totheir simple or quaternary salts.

Polymers of the above type include the following classes of polymers,and their derivatives: ethylene/maleic anhydride copolymers,isobutylene/maleic anhydride copolymer, 2-methyl-pentene-1/maleicanhydride copolymers, styrene/maleic anhydride copolymers,vinylacetate/maleic anhydride copolymers, a-methylstyrene/maleicanhydride copolymers, polymaleic anhydride polymers, polyacrylicanhydride polymers, polyacrylic acid polymers, octadecene-1/maleicanhydride copolymers, loweralkylaminoloweralkylimide ofoctadene-1/maleic anhydride copolymers, aliphatic ester ofethylene/maleic anhydride copolymers, vinylalkylether/maleic anhydridecopolymers, aliphatic methacrylate/methacrylamide copolymers, aliphaticmethacrylate/diloweralkylaminoloweralkyl methacrylate copolymers,loweralkylaminoloweralkylimide of styrene maleic anhydride copolymersand polymethacrylic acid polymers.

Individual examples of such polymers include ethylene/maleic anhydridecopolymer, the disodium salt of isobutylene/maleic anhydride copolymer,the calcium salt of styrene/maleic anhydride copolymer, hydrolyzedpropylene/maleic anhydride copolymer, the monopotassium salt ofdivinylether/maleic anhydride copolymer, hydrolyzed vinyl methylether/citraconic anhydride copolymer, half lauryl ester ofoctene-1/maleic anhydride copolymer, octadecene-1/maleic anhydridecopolymer, ethylene/maleic acid copolymer, the dipotassium salt ofisobutylene/maleic acid copolymer, the half amide half ammonium salt ofisobutylene/maleic anhydride copolymer, ethylene/acrylic acid copolymer,ethylene/acrylic anhydride copolymer, half capryl ester ofhexene-1/acrylic anhydride copolymer, ethylene/aconitic anhydridecopolymer, half ethylamide of styrene/maleic anhydric copolymer,ethylene/fumaric acid copolymer, octylamide acid of ethylene/maleicanhydride copolymer, octadecylamide ammonium salt ofvinylmethylether/maleic anhydride copolymer, dimethylaminopropylamideacid of divinylether/maleic anhydride copolymer, isobutylamide of vinylacetate/maleic anhydride copolymer, methiodide quaternary derivative ofN,N-dimethylaminoethylamide of polymaleic anhydride, octadecyl esterammonium salt of ethylene/itaconic anhydride copolymers, butylamine halfamide of hexene-1/chloromaleic anhydride copolymer, the partial diameterof ethylene/maleic anhydride copolymer, n-decylamide of decene-1/maleicanhydride copolymer, N,N-diethylaminopropylamide ammonium salt ofisobutylene/maleic anhydride copolymer, dimethyl sulfate quaternary saltof dimethylaminoethylamide of polymaleic anhydride, the partial halfhexylamide of vinylmethylether/maleic anhydride copolymer, thediammonium salt of ethylene/maleic anhydride copolymer, the monoamideacid of propylene/maleic anhydride copolymer, N-ethyl monoamide ofdivinylether/maleic anhydride copolymer, N-dodecyl monoamide ofvinylmethylether/maleic anhydride copolymer,N,N-dimethylaminopropylimide of triacontene/maleic anhydride copolymer,N,N-dimethylaminopropyl monoamide of styrene/citraconic anhydridecopolymer, n-butylmonoamide of polymaleic anhydride,N,N-diethylmonoamide ammonium salt of vinyl acetate/maleic anhydridecopolymer, n-butylimide of ethylene/maleic anhydride copolymer,octadecylimide of polymaleic anhydride, N,N-dimethylaminopropylimide ofstyrene/maleic anhydride copolymer, dimethylsulfate quaternary salt ofdiethylaminopropylimide of divinylether/maleic anhydride copolymer,N,N-dimethylaminopropyl half amide of paramethyl styrene/maleicanhydride copolymer, methyliodide quaternary salt of dimethylaminohexylhalf amide half ammonium salt of a-methylstyrene/maleic anhydridecopolymer, N,N-diethylaminoethyl half amide half sodium salt ofisobutylene/maleic anhydride copolymer, partial lauryl ester ofethylene/maleic anhydride copolymer, vinyl octadecyl ether/maleicanhydride copolymer, stearyl methacrylate/methacrylamide copolymer andstearyl methacrylate/N,N-dimethylaminoethyl methacrylate copolymer.

A particularly preferred class of polymers for use in the presentinvention are lipophilic polymers, that is, polymers which have alipophilic grouping or groupings included therein. A lipophilic groupingor moiety typically contains 6 or more atom units and may be in anysuitable form such as a polyalkylene or alkylene oxide containing 6 ormore atom units or as an ester, amide or imide unit containing 6 or moreatom units, for example, 6 to 30 carbon atoms formed by reaction of thecarboxyl containing monomer with lipophilic amines or alcohols such as,for example, hexanol, octanol, octylamine, hexylamine, octadecanol, etc.and the like. Examples of such preferred polymers include theoctadecylimide of polymaleic anhydride, the methyl iodide quaternarysalt of dimethylaminohexyl half amide half ammonium salt ofa-methylstyrene/maleic anhydride copolymer, dodecyl monoamide ofvinylmethylether/maleic anhydride copolymer, the octadecyl esterammonium salt of ethylene/itaconic anhydride copolymer, the decylamideof decene-1/maleic anhydride copolymer, octadecene-1/maleic anhydridecopolymer etc. and the like.

Pharmaceutically acceptable alkaline earth metals and alkali metals,such as calcium, magnesium and potassium are useful in preparingconveniently administered forms of the polyelectrolyte polymers of thisinvention. The salts of metals such as magnesium, aluminum, zinc, iron,barium and bismuth are also useful in the present invention.

Representative EMA-type carboxylic acid or anhydride-olefin polymers,especially maleic acid or anhydrideolefin polymers of the foregoing typeare known, for example, from U.S. Pat. Nos. 2,378,629; 2,396,785;3,157,595; and 3,340,680. Generally, the copolymers are prepared byreacting ethylene or other unsaturated monomer, or mixtures thereof,with the acid anhydride in the presence of a peroxide catalyst in analiphatic or aromatic hydrocarbon solvent for the monomers butnonsolvent for the interpolymer formed. Suitable solvents includebenzene, toluene, xylene, chlorinated benzene and the like. Whilebenzoyl peroxide is usually the preferred catalyst, other peroxides suchas acetyl peroxide, butyryl peroxide, ditertiary butyl peroxide, lauroylperoxide and the like, or any of the numerous azo catalysts, aresatisfactory since they are soluble in organic solvents. The copolymertypically contains from about 25 to about 75% (mole %) of the olefin andpreferably contains substantially equimolar quantities of the olefinresidue and the anhydride or acid residue; that is, a mole ratio ofolefin to anhydride or acid in the range of from about 2:3 to about 3:2.Generally, the copolymer will have a degree of polymerization of 8 to100,000 preferably about 100 to 5,000, and a molecular weight of about1,000 to 1,000,000, preferably about 10,000 to 500,000. The propertiesof the polymer, such as molecular weight, for example, are regulated byproper choice of the catalyst and control of one or more of thevariables such as ratio of reactants, temperature, and catalystconcentration or the addition of regulating chain transfer agents, suchas diisopropyl benzene, propionic acid, alkyl aldehydes, or the like.Numerous of these polymers are commercially available.

Derivatives containing basic or cationic groups can be prepared by anyconvenient procedure. Representative derivatives of polymers employed inthe present invention are known to the art, for example, from U.S. Pat.No. 3,398,092. One group of useful derivatives are those in which thecarboxyl groups are partially replaced with basic or cationic bearingmoieties. For example, useful derivatives are conveniently formed byreaction of the carboxyls with polyfunctional amines such asdimethylaminopropylamine or dialkylaminoalcohols such asdimethylaminoethanol, the former forming an amide linkage with thepolymer, or in certain cases at higher temperatures forming an imidelinkage with the vicinal carboxyls and the latter forming an esterlinkage. Such pendant free amine groups can then be converted, ifdesired, to their simple or quaternary salts.

Imides of a starting carboxyl or carboxylic acid anhydride containingpolymer, e.g., EMA, are produced by:

A. Heating a limiting amount of a secondary or tertiaryaminoloweralkylamine with the anhydride or carboxyl-containing form ofthe polymer in a suitable solvent (e.g. Xylene) at a temperature ofabout 140°-150° C. until water is no longer given off. Such a reactionsimultaneously results in formation of imide groups in proportion to theamount of amine added and in the reformation of anhydride groups for theremainder of the polymer units. In this manner, imide-polymer productsare formed which possess imide linkages, the remaining carboxyl groups,when present, being in the anhydride form.

B. Alternatively, a partial amide polymer product may be converted tothe partial imide polymer product by heating a partial amide-polymerproduct in vacuo at 140°-150° C. until water is no longer given off.Such an imide polymer product likewise possesses comparable proportionsof imide and anhydride groups depending upon the number of amide groupsoriginally contained in the starting partial amide-polymer product.

Partial secondary or tertiary aminoloweralkylamides of the startingcarboxyl or carboxylic acid anhydride-containing polymer, e.g., EMA, areobtained by contacting the polymer with a limiting amount of theselected amine in suspension in a solvent such as benzene or hexane,resulting in formation of a partial amide-acid-anhydride derivative ofthe polymer, or a corresponding amide-carboxylate derivative thereof.The number of amide groups is dependent upon the quantity of the amineused as compared with the quantity of polymer employed.

Partial aminoester-polymer products are most conveniently prepared byheating at reflux temperatures overnight a limiting quantity of theselected aminoalcohol and carboxyl or carboxylic acid anhydridecontaining polymer, e.g., EMA, in a dry organic solvent such as tolueneor dimethylformamide and with the optional use of an acidic or basiccatalyst such as p-toluenesulfonic acid or sodium alkoxide. Theresulting product contains ester groups, carboxylic acid groups andanhydride groups, the respective numbers of which are determined by thequantity of aminoalcohol used in the reaction compared to the amount ofpolymer employed and, in some cases, by the temperature at which thereaction is carried out.

Suitable blocking and unblocking of the amine moiety of the reactantemployed in preparing amides, esters or imides may be effected whenrequired. Residual, nonmodified, polymer units may optionally beconverted to neutral groups or units by attachment to the polymermolecule of compounds including alkylamines, aminoalcohols, andalcohols.

Alternatively, the cationic character of the polymer can be providedthrough incorporation of monomers which impart a basic or cationiccharacter such as C-vinyl pyridines, vinyl amine, the severalamino-substituted vinyl benzenes (or toluenes, etc.), amino-bearingacrylates (or methacrylates, etc.), vinyl imioazole, etc.

The invention will be understood more fully by reference to thefollowing specific examples. It is understood that the examples arepresented for the purpose of illustration only and are not intended as alimitation of the invention.

EXAMPLE 1

To determine the effect of polymers of the present invention in loweringblood cholesterol levels the following test was conducted.

A group of 144 Columbian male chicks (1 day old) was divided in 12 equalsub-groups. Each sub-group of 12 chicks was fed a basal diet and thesupplement shown in Table 1 for a 3-week period. Throughout the testperiod water was supplied ad libitum. The basal diet employed wasdesigned to raise the level of blood cholesterol in the chicks to whichit was fed. On conclusion of the feeding period the chicks weresacrificed and plasma cholesterol level was determined by the generalprocedure described in Chin. Chim. Acta. 10 381-84 (1964) by Levine andZak. Fecal cholesterol and fecal lipid were also determined. In the testthe copolymers of diets 2-10 and 12 are linear and the copolymer of diet11 is crosslinked. The results are set forth in Table 1:

    ______________________________________                                        Basal Diet                                                                    Ingredient           % by Weight                                              ______________________________________                                        Soybean Meal         25.00                                                    Whole Egg (Powder)   25.00                                                    Vitamin Supplement   0.40                                                     Choline Chloride     0.14                                                     Iodized Salt         0.05                                                     Delamix              0.10                                                     Dicalcium Phosphate  2.00                                                     Calcium Carbonate    1.00                                                     Vitamin A (10,000 μ/g)                                                                          0.10                                                     Vitamin D (1,500 μ/g)                                                                           0.008                                                    Alpha Cel            3.00                                                     Cerelose             42.752                                                   ______________________________________                                    

The results of this test show that the polyelectrolyte polymers of thepresent invention are effective in lowering blood cholesterol and inincreasing fat excretion.

                                      TABLE 1                                     __________________________________________________________________________                                              Plasma Fecal                                                          Level in Feed                                                                         Cholesterol                                                                          Cholesterol                                                                          Fecal Lipid           Diet                                                                             Polymer                        % by weight                                                                           mg/100 ml.                                                                           % dry weight                                                                         % dry                 __________________________________________________________________________                                                            weight                1. None (Control)                 --      213    0.75   4.70                  2. Octadecene-1/Maleic Anhydride Copolymer                                                                      0.5     183    1.06   6.9                   3. Octadecene-1/Maleic Anhydride Copolymer                                                                      1.0     150    1.32   9.28                  4. Octadecene-1/Maleic Anhydride Copolymer                                                                      2.0     125    1.38   7.74                  5. Octadecene-1/Maleic Anhydride Copolymer                                                                      3.0     107    1.43   10.29                 6. N,N-Dimethylaminopropyl-imide of Octadecene-1/Maleic                                                         3.0     114    1.58   24.7                     Anhydride Copolymer                                                        7. Partial Lauryl ester of ethylene/Maleic Anhydride Copolymer                                                  3.0     149    0.64   7.4                   8. Vinyloctadecyl ether/Maleic Anhydride Copolymer                                                              3.0     141    1.06   8.1                   9. Stearyl methacrylate/Methacrylamide Copolymer                                                                3.0     136    1.32   9.8                   10.                                                                              Stearyl methacrylate/N,N-dimethylamine ethyl methacrylate                                                    3.0     120    1.56   22.4                     copolymer                                                                     N,N-Dimethylaminopropylimide of styrene/maleic anhydride                                                     3.0     107    1.53   20.3                     copolymer                                                                     Octene-1/maleic Anhydride Copolymer                                                                          3.0     147    1.40   12.21                 __________________________________________________________________________

EXAMPLE 2

A group of 36 Vantress-Arbor Acre male chicks (1 day old) was dividedinto three equal subgroups. Each subgroup of 12 chicks was fed the BasalDiet of Example 1 and 3.0% of the supplement shown in Table 2 for a 3week period. Throughout the test period water was supplied ad libitum.On conclusion of the feeding period the chicks were sacrificed andplasma cholesterol level was determined by the procedure referred to inExample 1. In the test the copolymers of diet 14 and 15 werecrosslinked. The results are set forth in Table 2.

                  TABLE 2                                                         ______________________________________                                                                  PLASMA                                                                        CHOLESTEROL                                         DIET   POLYMER            mg/100 ml                                           ______________________________________                                        13     None               290.5                                               14     Ethylene/maleic anhydride                                                                        204.8                                               15     Ethylene/maleic anhydride                                                     hydrolyzed         192.8                                               ______________________________________                                    

EXAMPLE 3

The procedure of Example 2 was repeated. The polymers used and theresults obtained are given in Table 3.

                  TABLE 3                                                         ______________________________________                                                                  PLASMA                                                                        CHOLESTEROL                                         DIET   POLYMER            mg/100 ml                                           ______________________________________                                        16     None               309                                                 17     Ethylene/maleic anhydride                                                     (crosslinked)      275                                                 18     Ethylene/maleic anhydride                                                     calcium salt (crosslinked)                                                                       255                                                 ______________________________________                                    

EXAMPLE 4

The procedure of Example 1 is repeated in all essential details with theexception that the polymer is polymaleic anhydride to lower the bloodcholesterol level of the chicks.

EXAMPLE 5

The procedure of Example 1 is repeated in all essential details with theexception that the polymer is polyacrylic anhydride to lower the bloodcholesterol level of the chicks.

EXAMPLE 6

The procedure of Example 1 is repeated in all essential details with theexception that the polymer is polyacrylic acid to lower the bloodcholesterol of the chicks.

EXAMPLE 7

The procedure of Example 1 is repeated in all essential details with theexception that the polymer is polymethacrylic acid to lower the bloodcholesterol level of the chicks.

EXAMPLE 8

Cattle having a high level of blood cholesterol are fed a daily rationcontaining from 0.01 to 5.0% of octadecene-1/maleic anhydride and thelevel of their blood cholesterol is lowered.

EXAMPLE 9

Heavy weight Hampshire hogs having a high level of blood cholesterol arefed a daily ratio containing from 0.01 to 3.0% ofN,N-dimethylaminopropylimide of octadecene-1/maleic anhydride copolymerand the level of their blood cholesterol is lowered.

EXAMPLE 10

Aged chimpanzees having a high level of blood cholesterol are fed adaily ration containing from 0.05 to 1.0% of octadecene-1/maleicanhydride copolymer and the level of their blood cholesterol is lowered.

EXAMPLE 11

This example shows the effectiveness of octadecene-1/maleic anhydride(substantially equimolar copolymer) in the method of the presentinvention. Twenty-four New Zealand white rabbits weighing an average of2600 grams each were divided into three groups of equal number. Therabbits were fed ad libitum for 5 weeks a low cholesterol diet. Thefirst group (control) was fed Purina Rabbit Chow brand rabbit feedsupplemented with 2% animal tallow and 1% cellulose. The second groupwas fed the identical feed as the control with the exception that 1%octadecene-maleic anhydride was substituted for the 1% cellulose. Thethird group was fed the identical feed as the control with the exceptionthat 1% CHOLESTYRAMINE, a crosslinked divinyl-benzene-polystyrene ionexchange resin containing quaternary ammonium groups was substituted forthe 1% cellulose. On completion of the 5 week feeding period the animalswere sacrificed and the following measurements were made: (1) plasmacholesterol, (2) plasma triglyceride, (3) liver cholesterol, and (4)liver fat. The measurements were averaged and are presented in Table 4.These results when considered as a whole indicate that highera-olefin/maleic anhydride copolymers are effective in lowering bloodlipids, i.e., in the treatment of hypercholesterolemia.

EXAMPLE 12

This example shows the effectiveness of octadecene-1/maleic anhydride(substantially equimolar copolymer) in the method of the presentinvention. Forty New Zealand white rabbits weighing an average of 2600grams each were divided into five groups of equal number. The groupswere fed ad libitum for 5 weeks a high cholesterol diet. The first group(control) was fed Purina Rabbit Chow brand rabbit feed supplemented with2% animal tallow, 1% cellulose and 0.2% cholesterol. The second groupwas fed the identical feed as the control with the exception that 0.5%octadecene-maleic anhydride was substituted for 0.5% of the cellulose.The third group was fed the identical feed as the control with theexception that 1% octadecene maleic anhydride was substituted for the 1%cellulose. The fourth group was fed the identical feed as the controlwith the exception that 0.5% CHOLESTYRAMINE was substituted for 0.5% ofthe cellulose. The fifth group was fed the identical feed as the controlwith the exception that 1% CHOLESTYRAMINE was substituted for the 1%cellulose. Feces were collected during the final day of the test. Oncompletion of the 5-week feeding period the animals were sacrificed. Themeasurements performed in Example 11 above were carried out. Fecal fatwas also measured. The measurements were averaged and are presented inTable 5.

EXAMPLE 13

This example shows the effectiveness of polymers useful in the presentinvention. Four hundred and sixty-eight 1-day old chicks (Vantress-ArborAcre Cockerels) were divided into 13 groups of equal number. Each groupof chicks were fed ad libitum for 21 days a high cholesterol diet of soybean meal, whole egg powder, minerals and vitamins. Cholesterol loweringadditive in the amount indicated in Table 6 was added to the diet ofgroups 2-13. Group 1 served as a control. Feces were collected for a24-hour period between days 19 and 20. On completion of the 21-day testthe birds were sacrificed and the following measurements were made: (1)plasma cholesterol, (2) liver cholesterol, and (3) liver fat. Themeasurements were averaged and are presented in Table 6.

EXAMPLE 14

This example shows the effectiveness of representative members of (1) agroup of polymers which are copolymers of higher a-olefins (10-22 carbonatoms or more) and maleic acid or maleic anhydride and (2) a group ofpolymers which are copolymers of higher alkyl vinyl ethers (10-22 carbonatoms or more) and maleic acid or maleic anhydride. The maleic acidcopolymers were prepared by hydrolysis of the corresponding maleicanhydride copolymers in 5% acetic acid in water at 50° C for 24 hoursfollowed by freeze drying.

Thirty-six female rabbits (1.5-2.0 kilograms each) were placed inindividual cages. The rabbits were randomly divided into 12 groups (3animals per group) for feeding purposes and fed ad libitum for 3 weekshigh cholesterol diets. The first group (control) was fed a basal dietdesigned to induce hypercholesterolemia. The died consisted of PurinaRabbit Chow brand rabbit feed supplemented with 2% animal tallow, 0.2%cholesterol and 1% cellulose. The other 11 groups were fed the identicalbasal diet as the control group with the exception that 1% of acopolymer useful in the present invention was substituted for the 1%cellulose. On completion of the 3-week feeding period the animals wereweighed and blood samples were taken. The samples were analyzed forcholesterol (Technicon autoanalyzer). At the beginning of the test, ablood sample was taken from each animal (heart puncture) and analyzedfor cholesterol. All results are presented in Table 7. The effectivenessof a tested compound is shown by comparing the final plasma cholesterolof the animals treated with the compound with the final plasmacholesterol of the control animals.

EXAMPLE 15

Three baboons were fed a high cholesterol diet for 8 weeks. At thebeginning of the fourth week of feeding, octadecene-1/maleic anhydridecopolymer (0.5% by weight of the total diet) was added to the diet oftwo of the baboons and equal CHOLESTRYRAMINE, a crosslinkeddivinylbenzene-polystyrene ion exchange resin containing quaternaryammonium groups was added to the diet of the third baboon. In each ofthe three animals the blood cholesterol level was found to be lower atthe end of the eighth week than at the end of the third week with theblood cholesterol level at the end of the eighth week being lower in thetwo animals fed the diet containing octadecene/maleic anhydridecopolymer than in the animal fed the diet of CHOLESTRYRAMINE.

Representative formulations embodying polymers within the scope of thepresent invention are:

TABLET FORMULATION

The following formulation provides for the manufacture of 1,000 tablets:

    ______________________________________                                                                 GRAMS                                                ______________________________________                                        (1)  Octadecene-1/maleic anhydride copolymer                                                                 25                                             (2)  Lactose                   181                                            (3)  Corn Starch               92                                             (4)  Magnesium Stearate        2                                              ______________________________________                                    

Thoroughly granulate a mixture of 72 grams of cornstarch and the lactosewith a paste prepared by dissolving 20 grams of cornstarch in 100 ml ofhot distilled water. Dry the resulting granulation at 40°-45° C. andpass it through a No. 16-mesh screen. To the dried, screened granulationadd a blended mixture of the active ingredient (1) and the magnesiumstearate. Thoroughly blend and then press into tablets of 300 mg. each.

CAPSULE FORMULATION

The following formulation provides for the manufacture of 1,000capsules:

    ______________________________________                                                                 GRAMS                                                ______________________________________                                        (1)  N,N-dimethylaminopropylimide of                                               octadecene-1/maleic anhydride                                                 copolymer                 25                                             (2)  Lactose                   274                                            (3)  Magnesium Stearate        2                                              ______________________________________                                    

Mix active ingredient (1) with the lactose and blend in the magnesiumstearate. Fill hard gelatin capsules with 300 mg. each of the blendedmixture to produce capsules containing the active ingredient.

While the invention has been described with reference to particularembodiments thereof, it will be appreciated that modifications andvariations are possible without departing from the invention.

                                      TABLE 4                                     __________________________________________________________________________                     PLASMA       LIVER                                                            Cholesterol                                                                         Triglyceride                                                                         Cholesterol                                                                         Fat                                                        *mg % *mg %  % of wet weight                                 __________________________________________________________________________    Group 1                                                                            (Control)   52    82     .38   6.89                                      Group 2                                                                            (1 % Octadecene-                                                                          43    87     .36   5.12                                           maleic anhydride)                                                        Group 3                                                                            (1%         74    110    .39   5.69                                           CHOLESTYRAMINE)                                                          __________________________________________________________________________     *mg per 100 cc plasma.                                                   

                                      TABLE 5                                     __________________________________________________________________________                        PLASMA       LIVER    FECES                                                   Cholesterol                                                                         Triglyceride                                                                         Cholesterol                                                                         Fat                                                                              Fat                                                     *mg % *mg %  % of wet weight                                                                        % of dry wt.                        __________________________________________________________________________    Group 1                                                                            (Control)      672   106    .92   7.80                                                                             4.51                                Group 2                                                                            (0.5% Octadecene-maleic                                                                      186   113    .47   5.98                                                                             6.03                                     anhydride)                                                               Group 3                                                                            (1.0% Octadecene-maleic                                                                      89    56     .41   5.80                                                                             6.29                                     anhydride)                                                               Group 4                                                                            (0.5% CHOLESTYRAMINE)                                                                        356   68     .79   7.36                                                                             7.12                                Group 5                                                                            (1.0% CHOLESTYRAMINE)                                                                        310   105    .74   7.18                                                                             6.04                                __________________________________________________________________________     *mg per 100 cc of plasma.                                                

                                      TABLE 6                                     __________________________________________________________________________                               Feed  Plasma                                                                              Liver Liver Excreta                                                                             Excreta                                     Weight                                                                            Consumed                                                                            Cholesterol                                                                         Cholesterol                                                                         Fat   Cholesterol                                                                         Fat                                         Grams                                                                             Grams mg %  % wet wt.                                                                           % wet wt.                                                                           % dry                                                                               % dry                __________________________________________________________________________                                                             wt.                  Group 1                                                                             (control)        328 472   257   2.58  10.7  0.46  8.2                  Group 2                                                                             (Octadecene-maleic anhydride)                                                                  360 483   250   1.26  8.9   0.97  6.3                        (0.6%)                                                                  Group 3                                                                             (Octadecene-maleic anhydride)                                                                  338 509   236   0.57  7.6   1.03  10.1                       (1.2%)                                                                  Group 4                                                                             (CHOLESTYRAMINE) (0.6%)                                                                        364 480   181   0.50  7.6   1.09  11.9                 Group 5                                                                             (CHOLESTYRAMINE) (1.2%)                                                                        331 428   154   0.45  7.7   1.12  11.4                 Group 6                                                                             (Octadecene-maleic anhydride                                                                   354 491   226   0.89  8.2   0.83  11.2                       dimethylaminopropylimide-                                                     100% derivative) (0.6%)                                                 Group 7                                                                             (Octadecene-maleic anhydride                                                                   315 456   258   --    --    0.65  12.8                       dimethylaminepropylimide-                                                     67% derivative) (0.6%)                                                  Group 13                                                                            (Stearyl methacrylate/                                                                         369 455   274   --    --    0.84  9.5                        methacrylic acid) (0.6%)                                                Group 8                                                                             (Stearylmethacrylate/dimethyl-                                                                 233 395   206   0.57  7.2   1.01  17.3                       aminoethylmethacrylate)                                                       (0.6%)                                                                  Group 9                                                                             (Stearylmethacrylate/dimethyl-                                                                 370 501   262   --    --    0.80  14.1                       aminoethylmethacrylate)                                                       (0.6%)                                                                  Group 10                                                                            (Vinyloctadecyl ether/                                                                         361 463   242   0.87  7.9   1.11  12.0                       maleic anhydride) (0.6%)                                                Group 11                                                                            (Vinyloctadecyl ether/                                                                         360 514   298   --    --    0.97  9.5                        maleic anhydride) (0.6%)                                                Group 12                                                                            (Stearyl methacrylate/                                                                         358 439   282   --    --    0.73  9.5                        methacrylic acid) (0.6%)                                                Group 13                                                                            (Stearyl methacrylate/                                                                         369 455   274   --    --    0.84  9.5                        methacrylic acid) (0.6%)                                                __________________________________________________________________________

                                      TABLE 7                                     __________________________________________________________________________                      Weight Gain in Grams For                                                                    Plasma Cholesterol                                                                        Plasma Cholesterol                       Polymer    3 Week Period (Average)                                                                     Initial mg.%*(Average)                                                                    Final mg.%*                       __________________________________________________________________________                                                (Average)                         Group                                                                             1  (Control)  546           53          377                                   2  Octadecene/maleic                                                                        461           51          117                                      anhydride (2:3) (1)                                                        3  Octadecene/maleic                                                                        487           123         8.sup.(1)                                acid (2:3)                                                                 4  Octadecene/maleic                                                                         436.sup.(2)   147.sup.(2)                                                                                70.sup.(2)                             anhydride (1:1)                                                            5  Octadecene/maleic                                                                        288           61          112                                      acid (1:1)                                                                 6  Decene/maleic                                                                            228           67          153                                      anhydride (1:1)                                                            7  Decene/maleic acid                                                                       320           78          117                                      (1:1)                                                                      8  Octadecyl vinyl                                                                          460           91          162                                      ether/maleic anhy-                                                            dride                                                                      9  Octadecyl vinyl                                                                           279.sup.(3)   64.sup.(3)  344.sup.(3)                             ether/maleic acid                                                          10 Dodecyl vinyl                                                                            365           70          260                                      ether/maleic anhy-                                                            dride (1:1)                                                                11 Dodecyl vinyl                                                                            538           60          103                                      ether/maleic acid                                                             (1:1)                                                                      12 Hexene/maleic an-                                                                        670            60.sup.(4)  472.sup.(4)                             hydride (1:1)                                                          __________________________________________________________________________     *mg per 100 cc of plasma                                                      .sup.(1) mole ratio of moletin to anhydride                                   .sup.(2) animal died of diarrhea during test (average of 2 animals)           .sup.(3) animal suffered severe weight loss due to diarrhea during test       (average of 2 animals)                                                        .sup.(4) animal suffered severe weight loss due to diarrhea during test  

We claim:
 1. A method for lowering the blood cholesterol level in aliving animal in need thereof comprising orally administering to saidanimal an effective blood cholesterol lowering amount of a copolymer of(1) an alpha alkene of .[.about 8.]. .Iadd.10 .Iaddend.to about 22carbon atoms and (2) maleic acid or anhydride, or a pharmaceuticallyacceptable amide, ester, imide, salt thereof or mixtures thereof, saidcopolymer having a molecular weight in the range of from about 1000 to1,000,000.
 2. The method of claim 1 wherein the copolymer isoctadecene-1/maleic anhydride.
 3. The method of claim 1 wherein thecopolymer is decene-1/maleic anhydride.
 4. The method of claim 1 whereinthe copolymer is octadecene-1/maleic acid.
 5. The method of claim 1 inwhich the copolymer is a lipophilic copolymer of an alpha-alkene of 10to 22 carbon atoms and maleic acid or anhydride. .[.6. The method ofclaim 1 in which the copolymer is a polyelectrolyte copolymer of analkene of 10 to 22 carbon atoms..].
 7. The method of claim 1 in whichthe alkene has 10 to 18 carbon atoms.
 8. The method of claim 1 in whichthe maleic component of the copolymer is a mixture.
 9. The method ofclaim 1 in which the copolymer is a copolymer of maleic acid oranhydride.